1021.2.19第1版
2021.6.16改訂
2023.1.2 改訂

ファイザー社とモデルナ社のワクチンで使われているmRNA改造法
By D Weissman and K Karikó(人工の配列,詳細不明)
Cap(ヒトα-globinで代用)
stop

５’startCoding sequence

3’ UTR

5’ UTR

３’ Poly(A)binding
Poly-A tail protein

本来のRNA【３つのmRNAトランスフェクション法】
・TransIT™法
・リポフェクチン™法
・脂質ナノパーティクル法→これがワクチンで使われている

改造後のRNA

Cap: RNAの先頭を示すキャップ(帽子)構造

2’-O-methyltransferaseでcap1に改変

5’ UTR, 3’ UTR: 非翻訳領域？？

Coding sequence: 遺伝子情報

ウリジンをpseudouridine(Ψ)に置換 →免疫機構が回避でき、蛋白合成が2～10倍に

Poly-A tail: 尾部を示す200個前後のアデニン鎖 130個のアデニン鎖【改造RNAは体内で何をするのか？】・自然のRNAでは：ウイルス感染→OAS(酵素)活性化→2-5A(核酸分子)合成→RNase L活性化→RNA分解が進行
・改造RNAでは：
OASを活性化しない→コロナmRNAは分解されず細胞内に残る。マウス実験では１回の注射で中和抗体が9週まで増加。memory B cellsの消長は不明。T follicularhelper cells ↑ →germinal center ↑ →中和抗体↑。PKRを抑制。
(青字は,その後,別の研究者が発表したデータ)【改造RNAは逆転写されるか？】細胞内で逆転写が起こることが確認された【Spike蛋白のコーディング】

full-length S protein with deleted furin cleavage siteとした場合、CD4+ T cellの反応が最良【免疫反応】

CD4+, CD8+↑ → Th1 ↑ →IFN-γ ↑ (つまりTh1細胞優位のため再感染時にアレルギー反応は起さない)【改造の効果】

培養細胞実験で2倍長く残る。ラット実験12～78倍蛋白合成が増強。ヒトでの研究から60日間残ることが確認された

ワイズマン、カリコ両氏の名前が含まれた学術文献の一覧
全部で 32 編ありますが、そのすべてを参照した上で当サイトの執筆を行っています。
内容は多岐にわたり、かつ専門的です。個々の文献についてご質問があれば、メールにてお知らせください。
岡田正彦
1Zika virus protection by a single low-dose nucleoside-modified mRNA vaccination. Pardi

N, Karikó K, Weissman D, et al. Nature. 2017;543(7644):248-251.
2

Suppression of RNA recognition by Toll-like receptors: the impact of nucleosidemodification and the evolutionary origin of RNA. Karikó K, Weissman D, et al. Immunity.
2005;23(2):165-75.
3

In vitro transcription of long RNA containing modified nucleosides. Pardi N, Weissman D,Karikó K, et al. Methods Mol Biol. 2013;969:29-42.
4

A Single immunization with nucleoside-modified mRNA vaccines elicits strong cellularand humoral immune responses against SARS-CoV-2 in mice. Laczkó D, Karikó K, Weissman
D, et al. Immunity. 2020;53(4):724-732.e7.
5

Incorporation of pseudouridine into mRNA yields superior nonimmunogenic vector withincreased translational capacity and biological stability. Karikó K, Weissman D, et al. MolTher. 2008;16(11):1833-40.
6 Expression kinetics of nucleoside-modified mRNA delivered in lipid nanoparticles to miceby various routes. Pardi N, Kariko K, Weissman D, et al. J Control Release. 2015;217:345-51.
7 Nucleoside-modified mRNA vaccines induce potent T follicular helper and germinal
center B cell responses. Pardi N, Karikó K, Weissman D, et al. J Exp Med. 2018;215(6):15711588.
8mRNA: Fulfilling the promise of gene therapy. Weissman D, Karikó K. Mol Ther.

2015;23(9):1416-7.
9 Nucleoside-modified mRNA immunization elicits influenza virus hemagglutinin stalkspecific antibodies. Pardi N, Karikó K, Weissman D, et al. Nat Commun. 2018;9(1):3361.

10Generating the optimal mRNA for therapy: HPLC purification eliminates immune

activation and improves translation of nucleoside-modified, protein-encoding mRNA. Karikó
K, Weissman D, et al. Nucleic Acids Res. 2011;39(21):e142.
11 Characterization of HIV-1 Nucleoside-Modified mRNA Vaccines in Rabbits and Rhesus
Macaques. Pardi N, Karikó K, Weissman D, et al. Mol Ther Nucleic Acids. 2019;15:36-47.
12

HPLC purification of in vitro transcribed long RNA. Weissman D, Karikó K, et al.

Methods Mol Biol. 2013;969:43-54.
13 Incorporation of pseudouridine into mRNA enhances translation by diminishing PKR
activation. Anderson BR, Weissman D, Karikó K, et al, Nucleic Acids Res. 2010;38(17):588492.
14 Administration of nucleoside-modified mRNA encoding broadly neutralizing antibody
protects humanized mice from HIV-1 challenge. Pardi N, Karikó K, Weissman D, et al. NatCommun. 2017;8:14630.
15 Purification of mRNA Encoding Chimeric Antigen Receptor Is Critical for Generation ofa Robust T-Cell Response. Foster JB, Weissman D, Karikó K, et al, Hum Gene Ther.
2019;30(2):168-178.
16mRNA is an endogenous ligand for Toll-like receptor 3. Karikó K, Weissman D, et al. JBiol Chem. 2004;279(13):12542-50.
17 Increased erythropoiesis in mice injected with submicrogram quantities ofpseudouridine-containing mRNA encoding erythropoietin. Karikó K, Weissman D, et al. MolTher. 2012;20(5):948-53.
18 Naturally occurring nucleoside modifications suppress the immunostimulatory activityof RNA: implication for therapeutic RNA development. Karikó K, Weissman D. Curr Opin
Drug Discov Devel. 2007;10(5):523-32.
19 Nucleoside modifications in RNA limit activation of 2'-5'-oligoadenylate synthetase andincrease resistance to cleavage by RNase L. Anderson BR, Weissman D, Karikó K, et al.
Nucleic Acids Res. 2011;39(21):9329-38.
20 Inhibition of toll-like receptor and cytokine signaling--a unifying theme in ischemictolerance. Karikó K, Weissman D, et al, J Cereb Blood Flow Metab. 2004;24(11):1288-304.

21 Nucleofection induces transient eIF2α phosphorylation by GCN2 and PERK.
Anderson BR, Karikó K, Weissman D. Gene Ther. 2013;20(2):136-42.
22 Identification of Cyclobutane Pyrimidine Dimer-Responsive Genes Using UVBIrradiated Human Keratinocytes Transfected with In Vitro-Synthesized Photolyase mRNA.
Boros G, Weissman D, Karikó K, et al. PLoS One. 2015;10(6):e0131141.
23 Transfection of pseudouridine-modified mRNA encoding CPD-photolyase leads to repair
of DNA damage in human keratinocytes: a new approach with future therapeutic potential.
Boros G, Weissman D, Karikó K, et al. J Photochem Photobiol B. 2013;129:93-9.
24

gp340 expressed on human genital epithelia binds HIV-1 envelope protein and facilitatesviral transmission. Stoddard E, Karikó K, Weissman D, et al. J Immunol. 2007;179(5):312632.
25 Inhibition of HIV-1 infection by small interfering RNA-mediated RNA interference.
Capodici J, Karikó K, Weissman D. J Immunol. 2002;169(9):5196-201.
26 Exogenous siRNA mediates sequence-independent gene suppression by signaling
through toll-like receptor 3. Karikó K, Weissman D, et al. Cells Tissues Organs.
2004;177(3):132-8.
27

Short interfering RNA-mediated inhibition of herpes simplex virus type 1 gene

expression and function during infection of human keratinocytes. Bhuyan PK, Karikò K,Weissman D, et al. J Virol. 2004;78(19):10276-81.
28

Small interfering RNAs mediate sequence-independent gene suppression and induce

immune activation by signaling through toll-like receptor 3. Karikó K, Weissman D, et al. JImmunol. 2004;172(11):6545-9.
29 Cutting edge: innate immune system discriminates between RNA containing bacterial
versus eukaryotic structural features that prime for high-level IL-12 secretion by dendritic
cells. Koski GK, Karikó K, Weissman D, et al. J Immunol. 2004;172(7):3989-93.
30 Extracellular mRNA induces dendritic cell activation by stimulating tumor necrosis
factor-alpha secretion and signaling through a nucleotide receptor. Ni H, Karikó K,Weissman D, et al. J Biol Chem. 2002;12;277(15):12689-96.

31HIV gag mRNA transfection of dendritic cells (DC) delivers encoded antigen to MHC

class I and II molecules, causes DC maturation, and induces a potent human in vitro primary
immune response. Weissman D, Karikó K, et al. J Immunol. 2000;165(8):4710-7.
32 Nucleoside-modified VEGFC mRNA induces organ-specific lymphatic growth andreverses experimental lymphedema. Szőke D, Karikó K, Weissman D, et al. Nat Commun.
2021;12(1):3460.
＊著者名は、第１著者、およびワイズマンとカリコ両氏の 3 名のみとし、それ以外は「その他」を意味する et al.で略記しました。学術文献における著者名の順番は研究への貢献度を表すなど重要な意味を持ちますが、当サイトの主旨を考慮し、このような記述としました。これらの情報で文献の検索が可能であることを確認済みです。また Karikó 氏の「ó」を「o」と表記している文献もあり、そのまま記述しました。